PMID: 3756201Oct 3, 1986Paper

Solubilization and properties of Ca2+-dependent human platelet phospholipase A2

Biochimica Et Biophysica Acta
R M KramerD Deykin

Abstract

Using a sonicated dispersion of radiolabeled 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine as substrate, we found that phospholipase A2 activity of human platelets was enhanced 2.4-fold by albumin (1 mg/ml). The enzyme was recovered predominantly in the cytosolic fraction of platelets with less than a third of its activity being associated with the membrane fraction. In the presence of 24 mM n-octyl-beta-D-glucopyranoside (octylglucoside) phospholipase A2 was effectively (more than 90%) extracted from platelet lysates without solubilization of platelet membranes. Ion exchange chromatography of the soluble enzyme yielded a phospholipase A2 of unchanged total activity and great stability. This phospholipase A2 was active only in the presence of divalent cations (Ca2+ greater than Sr2+ greater than Mg2+ = 0), required albumin for optimal activity and exhibited exclusive positional specificity for the acyl ester bond at the 2-position of 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine. Indomethacin (500 microM), mepacrine (500 microM) and N-ethylmaleimide (4 mM) inhibited the phospholipase A2 by 69, 62 and 19%, respectively. The results are discussed in the light of previous findings on human platelet phospholipase A2.

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Citations

Jul 1, 1990·The Journal of Membrane Biology·R O Carlson, I B Levitan
Oct 1, 1987·Lipids·O M Filgueiras, F Possmayer
Oct 22, 2008·Cardiovascular Drugs and Therapy·John E Burke, Edward A Dennis
Jan 1, 1991·Pharmacology & Therapeutics·W L SmithD L DeWitt
Oct 1, 1995·Journal of Lipid Mediators and Cell Signalling·J D ClarkL L Lin
Aug 5, 1997·Proceedings of the National Academy of Sciences of the United States of America·J GabbetaA K Rao
Nov 18, 2008·Journal of Lipid Research·John E Burke, Edward A Dennis
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Jun 15, 1988·Thrombosis Research·F SekiyaY Saito
Feb 24, 2015·Journal of Molecular Recognition : JMR·Melanie Grandits, Chris Oostenbrink
Jan 9, 2013·Expert Opinion on Therapeutic Patents·Victoria Magrioti, George Kokotos
Apr 24, 1991·Biochimica Et Biophysica Acta·D K KimK Inoue
Apr 3, 1991·Biochimica Et Biophysica Acta·Y NozawaK Nagata
Jul 31, 1987·Biochemical and Biophysical Research Communications·S T Silk, M E DeMarco
Mar 1, 1989·Prostaglandins, Leukotrienes, and Essential Fatty Acids·A D Purdon, A K Rao
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Feb 28, 2006·The American Journal of Forensic Medicine and Pathology·Bl Meel
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Nov 3, 1993·Biochimica Et Biophysica Acta·I KudoK Inoue
Dec 4, 1990·Biochimica Et Biophysica Acta·C F BennettS T Crooke

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